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缺血再灌注损伤诱导心肌细胞自噬相关基因过表达
肖健1,何斌3,朱晓燕2,张宇峰1,康波1,王志农1*
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(1.第二军医大学长征医院胸心外科,上海 200003;3.上海交通大学附属新华医院麻醉科,上海 200092;2.第二军医大学基础部生理学教研室,上海 200433)
摘要:
目的探讨缺血再灌注(IR)损伤对心肌细胞自噬相关基因(autophagy-related gene,Atg)表达的影响。方法成年雄性SD大鼠随机分为2组:对照组(假手术组),于左前降支(left anterior descending, LAD)下方穿过5-0丝线,不结扎;缺血再灌注损伤组:结扎LAD,45 min后松结扎线再灌注120 min。利用RT-PCR方法,检测各组心肌Beclin 1和Atg5 mRNA表达;应用蛋白质印迹分析法检测心肌LC3蛋白表达变化。结果缺血再灌注损伤后心肌Beclin
关键词:  心肌再灌注损伤  心肌细胞  自噬
DOI:10.3724/SP.J.1008.2011.0776
投稿时间:2010-12-07修订日期:2011-06-22
基金项目:国家自然科学基金青年科学基金项目
Ischemia-reperfusion injury induced overexpression of autophagy-related genes in cardiomyocytes
XIAO Jian1,HE Bin3,ZHU Xiao-yan2,ZHANG Yu-feng1,KANG Bo1,WANG Zhi-nong1*
(1. Department of Cardiothoracic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China;3. Department of Anaesthesiology, Xinhua Hospital, Shanghai Jiaotong University, Shanghai 200092, China;2. Department of Physiology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China)
Abstract:
Objective:To compare the autophagy-related gene stimulated by ischemical reperfusion (IR) injury. Methods: The SD rats (250-300 g) were divided randomly into 2 group:A: (Control group) the rat underwent thoracotomy without left anterior descending coronary (LAD) ligated;B: (Ischemia-Reperfusion Injury group the left anterior descending coronary was ligated for 45min, followed by reperfusion for 2 h. The mRNA expression levels of Beclin1 and Atg5 were examined with RT-PCR method, and the protein of LC3 was determined with Western-blot method. Results: The relative amount of the mRNA expression level of Beclin1and Atg5 was enhanced by IR to 2.45 and 1.6 times, respectively. The LC3-Ⅱprotein increased, which increased the ratio of LC3-Ⅱ/LC3-Ⅰ(IR group, 1.4; Con group, 0.2). Conclusion: Ischemia-Reperfusion Injury may induce cardiomyocytes autophagy, which contribute to cell death, by increasing the expression of the Atg family.
Key words:  Ischemia-Reperfusion Injury,myocardiocyte,autophagy